Effect of High-Tunnel and Open-Field Production on the Yield, Cannabinoids, and Volatile Profiles in Industrial Hemp (Cannabis sativa L.) Inflorescence.

This study discovered the impact of high-tunnel (i.e., unheated greenhouse) and open-field production on two industrial hemp cultivars (SB1 and CJ2) over their yield parameters, cannabinoid development, and volatile profiles. Development of neutral cannabinoids (CBD, THC, and CBC), acidic cannabinoids (CBDA, THCA, and CBCA), and total cannabinoids during floral maturation were investigated. The volatile profiles of hemp flowers were holistically compared via HS-SPME-GC/MS. Findings indicated a high tunnel as an efficient practice for achieving greater total weight, stem number, and caliper, especially in the SB1 cultivar. Harvesting high-tunnel-grown SB1 cultivars during early flower maturation could obtain a high CBD yield while complying with THC regulations. Considering the volatile profiles, hemp flowers mainly consisted of mono- and sesquiterpenoids, as well as oxygenated mono- and sesquiterpenoids. Volatile analysis revealed the substantial impact of cultivars on the volatile profile compared to the production systems.

Milk-derived exosome nanovesicles: recent progress and daunting hurdles.

Raw milk is the foundation of quality and safety in the dairy industry, and improving milk source management is the fundamental guarantee. Milk-derived exosomes (MDEs) are nanoscale information transfer molecules secreted by mammary cells with unique content and high stability, which can be used not only as potential markers to analyze key traits of lactation, reproduction, nutrition and health of animals, but also help farm managers to take timely interventions to improve animal welfare, milk quality, and functional traits. Our review first outlines the latest advances in MDEs isolation and purification, compositional analysis and characterization tools. We then provide a comprehensive summary of recent applications of MDEs liquid biopsy in breed selection, disease prevention and control, and feeding management. Finally, we evaluate the impact of processing on the stability of MDEs to offer guidance for dairy production and storage. The limitations and challenges in the development and use of MDEs markers are also discussed. As a noninvasive marker with high sensitivity and specificity, the MDEs-mediated assay technology is expected to be a powerful tool for measuring cow health and raw milk quality, enabling dynamic and precise regulation of dairy cows and full traceability of raw milk.

Plant protein solubility: A challenge or insurmountable obstacle.

Currently, there is an increasing focus on comprehending the solubility of plant-based proteins, driven by the rising demand for animal-free food formulations. The solubility of proteins plays a crucial role in impacting other functional properties of proteins and food processing. Consequently, understanding protein solubility in a deeper sense may allow a better usage of plant proteins. Herein, we discussed the definition of protein solubility from both thermodynamic and colloidal perspectives. A range of factors affecting solubility of plant proteins are generalized, including intrinsic factors (amino acids composition, hydrophobicity), and extrinsic factors (pH, ionic strength, extraction and drying methods). Current methods to enhance solubility are outlined, including microwave, high intensity ultrasound, hydrostatic pressure, glycation, pH-shifting, enzymatic hydrolysis, enzymatic cross-linking, complexation and modulation of amino acids. We base the discussion on diverse modified methods of nitrogen solubility index available to determine and analyze protein solubility followed by addressing how other indigenous components affect the solubility of plant proteins. Some nonproteinaceous constituents in proteins such as carbohydrates and polyphenols may exert positive or negative impact on protein solubility. Appropriate protein extraction and modification methods that meet consumer and manufacturers requirements concerning nutritious and eco-friendly foods with lower cost should be investigated and further explored.

Nonenzymatic glycation as a tunable technique to modify plant proteins: A comprehensive review on reaction process, mechanism, conjugate structure, and functionality.

Plant proteins are expected to become a major protein source to replace currently used animal-derived proteins in the coming years. However, there are always challenges when using these proteins due to their low water solubility induced by the high molecular weight storage proteins. One approach to address this challenge is to modify proteins through Maillard glycation, which involves the reaction between proteins and carbohydrates. In this review, we discuss various chemical methods currently available for determining the indicators of the Maillard reaction in the early stage, including the graft degree of glycation and the available lysine or sugar, which are involved in the very beginning of the reaction. We also provide a detailed description of the most popular methods for determining graft sites and assessing different plant protein structures and functionalities upon non-enzymatic glycation. This review offers valuable insights for researchers and food scientists in order to develop plant-based protein ingredients with improved functionality.

Water-soluble fraction of pea protein isolate is critical for the functionality of protein-glucose conjugates obtained via wet-heating Maillard reaction.

Wet-heating Maillard reaction (MR) has been applied to improve the function of proteins by conjugating with soluble carbohydrates. However, the impact of soluble solutes particularly in plant protein on the degree of MR and the properties of the corresponding conjugates has yet to be discussed. In this study, high-intensity ultrasound (HIUS) was utilized to pretreat commercial pea protein isolate in order to improve its solubility. Two different fractions including soluble fraction (SUPPI) and whole solution (UPPI) of HIUS treated PPI were conjugated with glucose (G) to prepare SUPPI-G and UPPI-G, respectively, over a course of 24 h wet-heating at 80 °C. Conjugation was confirmed by the degree of glycation, SDS-PAGE, FTIR, and intrinsic fluorescence analysis. Color change and glucose content analysis showed that the degree of MR was greater when using SUPPI rather than UPPI. The solubility of SUPPI-G was further improved by 24 h of MR while it remained unchanged for UPPI-G. The emulsifying activity index and foaming capability of SUPPI-G were similar to those of UPPI-G. Interfacial properties determined by dynamic adsorption and dilatational rheology at both oil-water and air-water interface suggested that insoluble fraction of UPPI is essential to make stable emulsions and foams. In conclusion, the proportion of soluble protein in PPI is critical to its wet-heating MR based conjugation with glucose and the solubility of the conjugates.

Dairy products and constituents: a review of their effects on obesity and related metabolic diseases.

Obesity has become a global public health problem that seriously affects the quality of life. As an important part of human diet, dairy products contain a large number of nutrients that are essential for maintaining human health, such as proteins, peptides, lipids, vitamins, and minerals. A growing number of epidemiological investigations provide strong evidence on dairy interventions for weight loss in overweight/obese populations. Therefore, this paper outlines the relationship between the consumption of different dairy products and obesity and related metabolic diseases. In addition, we dive into the mechanisms related to the regulation of glucose and lipid metabolism by functional components in dairy products and the interaction with gut microbes. Lastly, the role of dairy products on obesity of children and adolescents is revisited. We conclude that whole dairy products exert more beneficial effect than single milk constituent on alleviating obesity and that dairy matrix has important implications for metabolic health.

Sensory profile of pulse-based high moisture meat analogs: A study on the complex effect of germination and extrusion processing.

Germination and extrusion are two processes that could affect beany flavors in pulse-based high-moisture meat analogs (HMMAs). This research studied the sensory profile of HMMAs made by protein-rich flours from germinated/ungerminated pea and lentil. Air-classified pulse protein-rich fractions were processed into HMMAs with twin screw extrusion cooking, optimized at 140 °C (zone 5 temperature) and 800 rpm screw speed. Overall, 30 volatile compounds were identified by Gas Chromatography-Mass Spectrometry/Olfactory. Chemometric analysis exhibited that the extrusion markedly (p < 0.05) reduced beany flavor. A synergistic effect of germination and extrusion process was observed, decreasing some beany flavors such as 1-octen-3-ol and 2,4-decadienal, and the overall beany taste. Pea-based HMMAs are suitable for lighter, softer poultry meat, while lentil-based HMMAs are suited for darker, harder livestock meat. Those findings offer novel insights into the regulation of beany flavors, odor notes, color, and taste to improve the sensory quality of HMMAs.

Unraveling the role of germination days on the aroma variations of roasted barley malts via gas chromatography-mass spectrometry based untargeted and targeted flavoromics.

Roasting imparts malts with an increased amount of hedonic aromas. However, the relationship between the production of roasted malts and the generation of characteristic malt aromas remains unclear. In this study, roasted barley malts (RM) were prepared from three consecutive germination days (3, 4, 5D), and the aroma profiles among RM and base malt were holistically compared via HS-SPME-GC-MS/O-based flavoromics. Furthermore, the wort color, free amino acids, reducing sugars, and fatty acids compositions were determined before-and-after roasting. Results showed that roasting could flatten variations of precursors regardless of germination days. Additionally, based on quantitation of 53 aromas, a PLS-DA model was applied to differentiate all malts by 17 aromas with VIP ≥ 1. As for aroma harmony, RM with 4D-germination outstood due to a pleasant nutty note with the highest sweet-to-nutty index of 0.8. This work answers how germination days would impact the aroma of RM for the first time.

Oleofoams stabilized by monoacylglycerides: Impact of chain length and concentration.

Oleofoams are plant oil based whipped systems which have drawn academic and industry attention in recent years. The aim of this study was to determine the effect of fatty acid chain length and monoacylglyceride (MAG) concentration on the performance and structural properties of MAG-based oleofoams. Four different MAGs (monolaurin, monomyrystin, monopalmitin, and monostearin) were studied at three concentration levels (5, 10, and 15 wt%). The fatty acid chain length had a statistically significant impact on the size and shape of crystals formed, while higher MAG concentrations led to higher numbers of crystals in the continuous oil phase. These differences affected the performance and physical properties of the oleofoams: compared to other MAGs, monostearin based oleofoams were harder and exhibited higher values of G' and G″, had higher overrun and showed better stability. Lastly, through microscopy techniques it was successfully proved that monostearin-based oleofoams are stabilized by both bulk and Pickering stabilization.

Effect of treatment of Fusarium head blight infected barley grains with hop essential oil nanoemulsion on the quality and safety of malted barley.

Fusarium mycotoxin contamination of malting barley has been a persistent food safety issue for malting companies. In this study, the effect of hop essential oil (HEO) nanoemulsion on fungal biomass and mycotoxin production during the malting process was evaluated. Furthermore, the localization of fungal hyphae on the surface and inside the tissue of barley and malts was observed. The application of HEO nanoemulsion reduced fungal biomass and deoxynivalenol (DON) contents at each stage of the malting process as compared to control. During malting process, the fungal hyphae on kernel surfaces was reduced appreciably after steeping. However, the increment of hyphae was observed between the husk and testa layer of barley after germination than raw barley grains. In addition to its antifungal activity, the antioxidant activity of HEO in the treated malts suppressed the formation of aldehydes. This study lays the foundation for the utilization of HEO in the malting industry.

Metal-Organic Materials (MOMs) Enhance Proteolytic Selectivity, Efficiency, and Reusability of Trypsin: A Time-Resolved Study on Proteolysis.

Proteases are involved in essential biological functions in nature and have become drug targets recently. In spite of the promising progress, two challenges, (i) the intrinsic instability and (ii) the difficulty in monitoring the catalytic process in real time, still hinder the further understanding and engineering of protease functionalities. These challenges are caused by the lack of proper materials/approaches to stabilize proteases and monitor proteolytic products (truncated polypeptides) in real time in a highly heterogeneous reaction mixture. This work combines metal-organic materials (MOMs), site-directed spin labeling-electron paramagnetic resonance (SDSL-EPR) spectroscopy, and mass spectrometry (MS) to overcome both barriers. A model protease, trypsin, which cleaves the peptide bonds at lysine or arginine residues, was immobilized on a Ca-MOM via aqueous-phase, one-pot cocrystallization, which allows for trypsin protection and ease of separation from its proteolytic products. Time-resolved EPR and MS were employed to monitor the populations, rotational motion, and sequences of the cleaved peptide truncations of a model protein substrate as the reaction proceeded. Our data suggest a significant (at least 5-10 times) enhancement in the catalytic efficiency (kcat/km) of trypsin@Ca-MOM and excellent reusability as compared to free trypsin in solution. Surprisingly, entrapping trypsin in Ca-MOMs results in cleavage site/region selectivity against the protein substrate, as compared to the near nonselective cleavage of all lysine and arginine residues of the substrate in solution. Remarkably, immobilizing trypsin allows for the separation and, thus, MS study on the sequences of truncated peptides in real time, leading to a time-resolved "movie" of trypsin proteolysis. This work demonstrates the use of MOMs and cocrystallization to enhance the selectivity, catalytic efficiency, and stability of trypsin, suggesting the possibility of tuning the catalytic performance of a general protease using MOMs.

Structural, functional properties, and volatile profile of hemp protein isolate as affected by extraction method: Alkaline extraction-isoelectric precipitation vs salt extraction.

The utilization of industrial hemp in food is gaining popularity. This study aimed to comprehensively investigate the impact of alkaline extraction-isoelectric precipitation (AE-IEP) and salt extraction (SE) dialysis on structural, functional properties and the volatile profile of hemp protein isolate (HPI). A higher protein content (97.21%) in SE extracted HPI (SE-HPI) was obtained than the AE-IEP extracted HPI (93.37%). In particular, protein subunit composition, structural properties were strongly influenced by the extraction methods. For example, SE-HPI exhibited a larger percentage of albumin (46.53%) and a lower amount of ß-sheet (52.65%) than its counterpart (albumin 20.92%, ß-sheet 54.46%). Consequently, SE-HPI showed the higher solubility, emulsion activity, and thermal stability. Interestingly, the volatile profile of the proteins showed that SE-HPI exhibited a lower number (21) of volatile compounds when compared to its counterpart (25). This study highlights the importance of establishing the relationships between extraction methods and functional attributes of HPI.

Lysozyme-phenolics bioconjugates with antioxidant and antibacterial bifunctionalities: Structural basis underlying the dual-function.

This work aims at adopting an Electron Paramagnetic Resonance (EPR) spectroscopic technique to help understanding protein-phenolic conjugation and final functionalities relationship as well as the underlying structural basis of antioxidant and antibacterial dual functionalities. Specifically, lysozyme (Lys) was conjugated with two natural phenolic acids, i.e. rosmarinic acid (RA) and gentisic acid (GA, our previous work) with obviously different molecular features. Lys-RA displayed 8.6- and 4.0-times enhanced antioxidant stoichiometry compared to the native Lys and ones with GA, respectively, due to the stronger antioxidant activity of RA. However, RA conjugation mitigated both enzymatic and antibacterial activities of Lys-RA conjugates. Such inhibition effect is attributed to the greater structural and surface property changes of Lys upon conjugating with RA. Furthermore, the polyphenol conjugation related structural basis of disturbance, reactivity and selectivity were explored via site-directed spin labeling (SDSL)-EPR. A dynamic picture of reactivity and selectivity of phenolics conjugation on Lys was proposed.

Antifungal activity, mycotoxin inhibitory efficacy, and mode of action of hop essential oil nanoemulsion against Fusarium graminearum.

This work aims to investigate antifungal, mycotoxin inhibitory efficacy of the hop essential oil (HEO) nanoemulsion and their mode of action (MOA) against Fusarium graminearum isolate, a fungal pathogen causing Fusarium Head Blight (FHB) in cereal crops. The HEO, primarily consisting of terpenes and terpenoids, was encapsulated in nanoemulsion droplets. Physically stable HEO-in-water nanoemulsion was fabricated using 0.5 wt% of tween 80 and 5 wt% oil phase comprising 30 % of Ostwald ripening inhibitor and 70 % of HEO. In terms of antifungal effect, HEO nanoemulsion could not only effectively inhibit mycelial growth and spore germination of F. graminearum isolates, but also remarkably suppress the production of deoxynivalenol (DON) and its derivatives in rice culture by applying 750 µg of HEO/g rice. Our studies on the MOA showed that HEO nanoemulsion could alter the contents of total lipid and chitin in outer cell membrane as well as damaging cytoplasmic membrane.

Molecular interaction mechanism and structure-activity relationships of protein-polyphenol complexes revealed by side-directed spin labeling-electron paramagnetic resonance (SDSL-EPR) spectroscopy.

Deciphering interactions between bioactive protein and polyphenols are critical for designing and controlling functional protein-polyphenol complexes. Herein, using the site-directed spin labeled T4 lysozyme (T4L) and rosmarinic acid (RA) as a model system, we combined electron paramagnetic resonance spectra to investigate molecular interaction mechanism of the protein-polyphenol complexes in structural or conformational details. Experimental results show that molecular interactions between T4L and RA are a process from order to disorder. TEM images display that the complexes finally assemble into quasi-spherical colloidal particles. When T4L/RA ratio is 1:1, the complexes exhibit the optimized enzymatic and antioxidant dual-functionalities due to the synergetic effect and protection mechanism. However, with excess addition of RA, the enzymatic and antioxidant activities of the complexes started to attenuate because the catalytic active site and bioactive hydroxyl groups were buried. The revealed high-resolution interaction process could help better understand the corresponding alterations between structure and functionalities.

Expanding the "Library" of Metal-Organic Frameworks for Enzyme Biomineralization.

Metal-organic frameworks (MOFs) are advanced platforms for enzyme immobilization. Enzymes can be entrapped via either diffusion (into pre-formed MOFs) or co-crystallization. Enzyme co-crystallization with specific metals/ligands in the aqueous phase, also known as biomineralization, minimizes the enzyme loss compared to organic phase co-crystallization, removes the size limitation on enzymes and substrates, and can potentially broaden the application of enzyme@MOF composites. However, not all enzymes are stable/functional in the presence of excess metal ions and/or ligands currently available for co-crystallization. Furthermore, most current biomineralization-based MOFs have limited (acid) pH stability, making it necessary to explore other metal-ligand combinations that can also immobilize enzymes. Here, we report our discovery on the combination of five metal ions and two ligands that can form biocomposites with two model enzymes differing in size and hydrophobicity in the aqueous phase under ambient conditions. Surprisingly, most of the formed composites are single- or multiphase crystals, even though the reaction phase is aqueous, with the rest as amorphous powders. All 20 enzyme@MOF composites showed good to excellent reusability and were stable under weakly acidic pH values. The stability under weakly basic conditions depended upon the selection of enzyme and metal-ligand combinations, yet for both enzymes, 3-4 MOFs offered decent stability under basic conditions. This work initiates the expansion of the current "library" of metal-ligand selection for encapsulating/biomineralizing large enzymes/enzyme clusters, leading to customized encapsulation of enzymes according to enzyme stability, functionality, and optimal pH.

Structural, and functional properties of phosphorylated pea protein isolate by simplified co-spray drying process.

In this work, to improve the functionality of pea protein isolate (PPI), sodium hexametaphosphate (SHMP) was added during last step of protein extraction and co-spray dried. The influence of PPI to SHMP mixing ratios (95:5 and 90:10) and reaction pH conditions (pH 6, 7, 8, and 9) on reaction efficiency, structural and functional properties of phosphorylated PPI were evaluated. Results showed that both mixing ratios had a similar degree of phosphorylation, suggesting the high efficiency of a 95:5 mixing ratio. The mixing ratio affected powder yield and proximate composition whereas hydrophobicity and denaturation temperature were regulated by pH conditions. For functionality, both mixing ratios showed significantly increased solubility at pH 6. Moreover, an increase in foaming capacity was observed in all phosphorylated PPI. The result from the current study may work as a basis for PPI phosphorylation in the food industry using the simplified method.

Uncovering aroma boundary compositions of barley malts by untargeted and targeted flavoromics with HS-SPME-GC-MS/olfactometry.

In this study, HS-SPME/GC-MS based untargeted and targeted flavoromics combing with olfactometry were employed to uncover aroma boundary compositions of five types of commercial barley malts with a wide range of Lovibond (L), including kilned base malts (1.8 L and 3.5 L) and roasted caramel malts (10 L, 60 L, and 120 L). Thirty-two compounds were identified as aroma-active with modified detection frequency (MF) > 50%. 3-Methylbutanal (malty), (2E)-nonenal (fatty, cardboard-like), and 2-furfural (burnt, bready) were recognized as the most influential odorants with MF > 70% in all the malts. After untargeted flavoromics, twenty-eight aromas were retained and quantitated. Furthermore, aroma boundary compositions inside/among malt groups were explored with PLS-DA. Eight aroma markers, 3-methylbutanal, 2-isopropyl-5-methyl-2-hexenal, (2E,4E)-Decadienal, 2-furfual, maltol, 2-acetylpyrrole, phenylacetaldehyde, and ethyl hexadecanoate were shortlisted for aroma boundary compositions regarding to the Lovibond of malts.

Structural characteristics of pea protein isolate (PPI) modified by high-pressure homogenization and its relation to the packaging properties of PPI edible film.

This study modified pea protein isolate (PPI) structure by high-pressure homogenization (HPH) and investigated PPI structural relation to the packaging properties of PPI film. HPH decreased PPI particle size, reduced surface charge, increased surface hydrophobicity, and increased free sulfhydryl, providing greater potential for covalent bonding during film formation. HPH decreased opacity of PPI films from 7.39 to 4.82 at pressure of 240 MPa with more homogeneous surface. The tensile strength and elongation at break were increased from 0.76 MPa to 1.33 MPa and from 96% to 197%, respectively, after treatment at 240 MPa. This improvement was due to the enhanced protein-protein and protein-glycerol hydrogen bonding as evidenced by FTIR. Increased ß-sheet and decreased α-helix by HPH was also observed, and ß-sheet was highly correlated to film tensile strength (Pearson coefficient of 0.973, P < 0.01). Principle component analysis visualized the influence of HPH treatment, and confirmed the association between structural characteristics and film properties.

Modification of physicochemical, functional properties, and digestibility of macronutrients in common bean (Phaseolus vulgaris L.) flours by different thermally treated whole seeds.

The utilization of beans (Phaseolus vulgaris L.) is hindered by unpleasant flavors, low macronutrients digestibility, and long cooking time. The pre-thermally treated beans can overcome these limitations. Therefore, the effect of thermal methods (moist-heat and dry-heat) and bean market classes (black, navy, kidney, and pinto) on functional properties and digestibility of bean flours were compared to raw counterparts. Within bean class, moist-heated samples showed increased water-holding capacities of 2.54-2.87 g H2O/g sample and starch/protein digestibility whereas dry-heated samples showed enhanced flavor profile and increased oil-holding capacities of 1.04-1.14 g oil/g sample. Among bean classes, moist-heated kidney bean flour showed the highest water-holding capacity of 2.87 g H2O/g sample and starch/protein digestibility while dry-heated pinto bean flour had the highest oil-holding capacity of 1.14 g oil/g sample. The current result may provide a basis for the development of pre-thermally treated legume flours and facilitate their applications.